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Microscopy

Stockroom Recipes
These recipes were collected over the course of many
years by various stockroom technicians (therefore I can take no
credit or responsibility for their accuracy or lack thereof).
For each recipe there are surely other ways of making them and/or
other formulations. Some are actually formulas concocted by our
faculty for their own research needs.
There are many duplications in this list (ie. rat ringers is also
listed as ringers, rat etc.) as I have intended it to be easily
downloaded and printed to a card type file with cross references.
This is most easily done by "searching" for the ******* lines
and "replacing" with a page break. The resulting pages should
fit easily on a 4x6 cards.
Should you wish to add or correct errors in this list
please e-mail your recipe or correction to me, Bob McNulty
and I will incorporate it/them into our list with the appropriate
acknowledgements (if desired).

BEFORE MIXING ANY REAGENT
YOU SHOULD UNDERSTAND THE
PROPERTIES OF ALL OF THE
CHEMICALS INVOLVED.
Consult your MSDS, Merk Index, CRC etc. before mixing any reagent.
The Biology Department, California State University Chico,
and most important, Myself.... assume NO responsibility for
the outcome of mixing any of these reagents !!!!!
Now on with the list

A's
2-4 - Dichlorophenoxyacetic acid (2-4-D)
Mix dry reagent in a small amount of 95% ethanol.
Place this solution in a warming plate 32 deg C and add
prewarmed water slowly with swirling until desired
concentration is reached.
****************************************************************
5% Ammoniacal Silver Nitrate
Add concentrated ammonium hydroxide to 5% silver nitrate
until resulting precipitate again dissolves.
****************************************************************
A.F.A. , Botanical
115 ml
Formalin
115 ml
Glacial acetic acid
805 ml
95% Ethanol
805 ml
Distilled water
****************************************************************
ACD used for blood
1.32 gm
Na Citrate
0.48 gm
Citric Acid
1.47 gm
Dextrose
100 mls
Distilled water
Use 0.25 ml/ml of blood
****************************************************************
ACETIC ACID - DILUTIONS
172 MLS
3N
57.3 mls
1N
1.046 mls
0.02N
Bring to a final volume of 1 Liter.
****************************************************************
Aceto Carmine Stain ( Stern)
45 ml
Glacial acetic acid
55 ml
water
0.5 gm
Carmine
Heat the 45% acetic acid solution to boiling. Add the 0.5gm
Carmine and continue heating for several minutes with
stirring. Filter and cool.
****************************************************************

ACETO-CARMINE (BELLING'S)
STOCK 1
55 ml
45 ml
5.0 gms
STOCK 2
55 ml
45 ml
5 gms

Distilled water
Acetic acid
Carmine

Distilled water
Acetic Acid
Ferric oxide
(iron acetate)
Boil stock solution 1 for 15 minutes, cool and filter
To 50 mls of stock 1 add stock 2 until pcpt just appears
then quickly add 50 ml more of stock 1
***for most purposes omit stock solution 2 and just use
stock solution 1.
****************************************************************
ACETO-CARMINE (chromosome stain for drosophila)
1. Weigh sufficient solid Carmin or Orcein, or Lacmoid to
make a 2% solution.
2. Place the stain with the appropriate volume of 45%
acetic acid in a flask with a vertical condensor.
3. Reflux all day, glass beads or pumice will help to
prevent bumping.
4. Allow to cool and filter.
****************************************************************
ACETOCARMINE
Stock sol'n - Boil and excess (aprox 0.5g/100 ml) of carmine
in 45% acetic acid (aq) 2-4 minutes. Cool and filter.
Working sol'n. Dilute stock sol'n with 45% acetic acid 1:2.
An iron mordanted stain often is favored because of darker
bluish tinged red. Belling added a few drops of 50% ferric
hydrate in 50% acetic acid, but only a few drops!
Too much iron will produce a pcpt in a short time.
Moree(1944) determined quantatively the amount of ferric
chloride to add and includes volumes tables
Moree, r. 1944 Stain tech 9:103-108
****************************************************************

ACETIC ALCOHOL
100 mls
Ethanol anhydrous
33 ml
Glacial Acetic Acid
****************************************************************
ACID ALCOHOL
50 ml
35% Ethanol
3 drops
HCl (con)
****************************************************************
ACID ALCOHOL FOR GLASSWARE CLEANING
100ml
70% Ethanol
5 ml
HCl
****************************************************************
ACID CLEANING SOLUTION
120 gms
Na Dichromate
1 Liter
Tap Water
1.6 Liters
Con Sulfuric
Dissolve chromate in water. Add acid with swirling. Be
sure to surround mixing vessel with ice bath.
****************************************************************
Acid cleaning solution 1
120 gm
Na dichromate
1000 ml
tap water
1600 ml
sulfuric acid
Dissolve chromate in water. Add acid with swirling. Be
sure to surround mixing vessel with ice bath.
****************************************************************
ACID CLEANING SOLUTION 2
120 gm
Potassium Dichromate
600 mls
water
920 mls
Con Sulfuric Acid
Dissolve chromate in water. Add acid with swirling. Be
sure to surround mixing vessel with ice bath.
****************************************************************
Acid cleaning solution 2
120 gm
Potassium dichromate
600 ml
water
920 ml
sulfuric acid
Dissolve chromate in water. Add acid with swirling. Be
sure to surround mixing vessel with ice bath.
****************************************************************

add more water and boil it down until only 90 mls remain. **************************************************************** Alum Cochineal 6 gm Potassium alum 6 gm powdered cochineal 90 ml distilled water Boil for 1/2 hour. Then add 70% ethanol (100 ml) and allow to stand for a day or two then filter. **************************************************************** ALEXANDER'S AFA 460 ML FORMALIN 1152 ML 95% ETHANOL 76 ML GLACIAL ACETIC ACID 2308 ML TAP WATER **************************************************************** ALGAL PRESERVATIVE 8 parts 50% Ethanol 1 part 5% Formalin 1 part Glycerine **************************************************************** Alizerine Red S for whole mount staining of bone 1 part alizerin red in 10. Cool and filter. After the fluid has settled. decant the supernatant liquid.Iodine NaOH 500 gm or KOH 700 gm *******and********** NaI 135 gm or KI 150 gm dissolved in distilled water to make 1 Liter (used for determining dissolved oxygen) **************************************************************** Alkaline Alcohol Add a few drops of 1.2% KOH Material is cleared in 2% KOH first.AFA (ALEXANDER'S) 460 ML FORMALIN 1152 ML 95% ETHANOL 76 ML GLACIAL ACETIC ACID 2308 ML TAP WATER **************************************************************** ALCOHOLIC BORAX CARMINE ( GRENACHER'S) 3 gm Carmine 4 gm Borax 100 ml distilled water Boil until the carmine is dissolved (1/2 hr) or better yet allow the mixture to stand at room temp until dissolved. Filter when cool and add a bit of thymol or a little salicylic acid to prevent the growth of mold **************************************************************** Aman's Lactophenol 20 ml melted phenol 20 ml Lactic acid 40 ml glycerine 21 ml distilled water To 100 ml of the above add 5 ml of 1% aqueous Cotton Blue or Acid Fuchsin.000 parts 1% .0% sodium bicarbonate to 70% Ethanol **************************************************************** Alkaline Na I Dissolve 40 gm of NaOH and 90 gm of NaI to make 100 ml of solution using distilled water. **************************************************************** Alum Carmine (Grenacher's) 100 ml Distilled water 3 gm Aluminum ammonium sulfate 1 gm Carmine Combine and boil for 15 mon or until carmine is dissolved. **************************************************************** . **************************************************************** Alkaline .

cool and filter To 50 mls of stock 1 add stock 2 until pcpt just appears then quickly add 50 ml more of stock 1 ***for most purposes omit stock solution 2 and just use stock solution 1.3% creatin in 40% potassium hydroxide but ADD JUST BEFORE USE.12 ml of 50% acetic acid. Keep in brown tightly stoppered bottle. **************************************************************** Baritt's Reagent test for acetylmethylcarbinol 5% solution of alpha naphthol in absolute ethyl alcohol.P. **************************************************************** Benzidine Reagent Make a saturated solution of benzidine in glacial acetic acid.5 gm of 1-amino-2-naphthal-4-sulfonic acid with 5 ml of sodium sulfite solution containing 1 gm anhydrous sodium sulfite per 5 ml and disolve in 200 ml sodium bisulfite solution containting 30 gm Na2S2O5 per 200 ml.3 gm Copper Sulfate to make 1 liter Distilled water Dissolve sodium citrate and sodium carbonate in 800 ml of distilled water. 0. **************************************************************** BELLING'S ACETO . add 0.8 gm Potassium oxalate 100 ml distilled water **************************************************************** Aqua Regia 1 part Nitric Acid 3 parts HCl 1 part distilled water WARNING!!! IF YOU DON'S ADD THE WATER POISONOUS CHLORINE GASS WILL FORM!!!!!!!!!! **************************************************************** B's Balsam in 100% Ethanol 1 part Balsam (solid) 1 part absolute ethanol This takes at least 3 days to disolve so prepare in advance. grade (28%) = 15 N **************************************************************** Ammonium Potassium Oxalate 1. **************************************************************** Barfoed's Solution Place 4. 40% solution of potassium hydroxide. Stable for 2 weeks.5 gm crystalized neutral cupric acetate in 100 ml of water.CARMINE STOCK 1 55 ml 45 ml 5. **************************************************************** Baryta Water Add Barium Hydroxide plus a pinch of Barium Chloride to a bottle fo distilled water until no more will go into solution. Filter and add copper sulfate dissolved in 100 ml of distilled water.S. Filter and keep tightly stoppered. Add distilled water to make 1 Liter. **************************************************************** Ammonium Hydroxide Concentrations Reagent grade (58%) = 29.7 N U. **************************************************************** Benedict's Reagent 173 gm Sodium Citrate 100 gm Sodium Carbonate (anhydrous) 17.2 gm Ammonium oxalate 0. .0 gms STOCK 2 55 ml 45 ml 5 gms Distilled water Acetic acid Carmine Distilled water Acetic Acid Ferric oxide (iron acetate) Boil stock solution 1 for 15 minutes.Aminonaphthal Sulfonic Acid Grind 0.

**************************************************************** Berlese's Medium 10 ml water 3 ml glacial acetic acid 5 ml dextrose syrup 8 gm gum acacia 75 gm Chloral Hydrate Mix water. Stir at intervals. Then add this to the phosphoric acid. and cut into strips for use in tests for proteins.Benzidine Test Place a drop of blood in 3 ml of water. cool covered. and syrup. Bottle in PLASTIC BOTTLES. Dissolve gum acacia in this mixture (may take a week). Add the 600 mls of 10% NaOH to the cooled water. Be careful not to include any air bubbles. mix and filter. Now add 500 mls of water. Boil 800 mls of distilled water. **************************************************************** .dd chloral hydrate. Then add 70% ethanol (100 ml) and allow to stand for a day or two then filter. then dissolve potassium sodium tartrate in this solution. **************************************************************** Biuret Reagent and Paper Biuret Reagent is prepared by adding 25 ml of 3% Copper Sulfate in 10% Potassium hydroxide.3 ml Glacial acetic acid 200 ml distilled water **************************************************************** Borax Carmine. Afeter gum acacia is dissolved. a. **************************************************************** Biuret Reagent tried and true!!!! 3 gm CuSO4*5H2O 12 gm Potassium sodium tartrate 600 ml 10% NaOH (aq) Dissolve copper sulfate in 500 ml of distilled water. **************************************************************** Botanical F. **************************************************************** Blood Diluting Fluid (Gower's) 12. acid. Grenacher's 3 gm Carmine 4 gm Borax 100 ml distilled water Boil until the carmine is dissolved (1/2 hr) or better yet allow the mixture to stand at room temp until dissolved. To this add 1 ml of 3% hydrogen peroxide. 115 ml Formalin 115 ml Glacial acetic acid 805 ml 95% Ethanol 805 ml Distilled water **************************************************************** Bouin's Fixative (Alcoholic) Kistner 150 ml 90% Ethanol 60 ml Formalin 15 ml Glacial acetic acid 1 gm Picric Acid **************************************************************** Bouin's Picro-Formol Fixative 75 parts saturated aq Picric Acid 25 parts Formalin 5 parts Glacial acetic Acid 1 gm of picric acid will saturate about 75 mls of water. Add and mix the remaining 350 mls of water bottle and store in refrigerator. **************************************************************** Bradford Protein Assay Dye Reagent 50 mg Coomassie Blue G250 100 ml 85% H3PO4 850 ml H2O 50 ml methyl alcohol Mix commassie blue in methyl alcohol first. with stirring to a 40% solution of NaOH unitl the mixture becomes a deep blue color. Then filter paper may be immersed in the reagent. Alchohlic. Mix the two solutions and bring to 2 liters. Add 2 ml of saturated solution of benzidine in glacial acetic acid. Paper (Walkers modification) Add 1% copper sulfate solution a drop at a time.A.A.5 gm Sodium Sulfate 33. dried.

2M Acid Potassium phosphate add the indicated number of ml of 0.2M KCl add the indicated number of mls of a 0.4 gm Brom Thymol Blue 500 ml 95% Ethanol 500 ml Distilled water Dissolve the indicator in the alcohol. Add 2 g of Charcoal. **************************************************************** .2M Potassium acid bi phthalate add the indicated number of mls of 0.2N Boric acid in 0.47gm /1000ml water of Na2-HPO4 Combine the two stock solutions as indicated **************************************************************** Buffer. stir and then filter through fast paper. Mix 40 ml of this stock solution with the amount of 1N NaOH indicated below and dillute to a final volume of 100 ml using water.Group 3 To 50 ml of a 0. and Phenol red.indicator 0.1 gm Brom Thymol Blue 16.5 ml of ). INDICATORS: Brom thymom blue. **************************************************************** Brom Thymol Blue .08gm /1000ml water of KH2-PO4 b) = 9. Dillute to 100 ml (Enough additional to completely disolve the reagent) **************************************************************** Brom Thymol Blue .3 gm of tris-hydroxymethyl aminomethane in 500 ml of water. Tris-Maleate Dissolve 29 gm of maleic acid and 30.2M Potassium acid bi phthalate add the add the indicated number of mls of 0. Brom phenol Blue.6 gm Brom Cresol Purple Mix BCP in dry morter with 18. INDICATORS: Cresol red and thymol blue **************************************************************** Buffer.Brazil's Fixative 150 ml 1 gm 60 ml 15 ml 80% Ethanol Picric Acid Formalin Acetic acid ADD JUST BEFORE USE **************************************************************** Brodie's Manometer Fluid 23 gm NaCl 5 gm Sodium Choleate 500 ml distilled water density = 1.indicator 0. **************************************************************** Brom Cresol Purple .033.Group 5 To 50 ml of 0. but some of these tend to decompose in the manometer.Group 1 To 50 ml of a 0. and dilute with distilled water to 200 ml. Add the distilled water. INDICATOR: Thymol Blue Buffer Solutions of Clark and Lubs .Group 2 To 50 ml of a 0. Determine the density of the solution with a pycnometer.Group 4 To 50 ml of a 0. Po = 10000 Evans blue (200mg/L) or acid fuchsin are excellent dyes for the fluid. shake and let stand for 10 min.2N HCl solution and then diltue to 200mls with distilled water. INDICATORS: Brom cresol green.1N NaOH.2N NaOH and dillute to 200 ml. Phosphate.0 ml 0.2N HCl and dilute to 200 ml with distilled water INDICATOR: Thymol blue and Brom Phenol blue Buffer Solutions of Clark and Lubs . other dyes may be used. Filter.2N NaOH. Use 5 drops / 10 ml of solution being checked. Buffer Solutions of Clark and Lubs . Sorensen's Stock solution a) = 9. Brom cresol purple Buffer Solutions of Clark and Lubs .2M KCl add the indicated number of ml of 0.2N NaOH and diltue to 200 ml with distilled water. **************************************************************** Budding Yeast Place a small portion of a yeast cake in malt extract broth Incubate while shaking. Brom cresol blue.indicator for fermentation 1.01N NaOH Mix in a mortar and then dillute to 250 ml. **************************************************************** Buffer Solutions of Clark and Lubs .

Pour dye solution into phenol solution (DO NOT REVERSE THIS! Let stand overnight. **************************************************************** Carmine. the soltuion of inom alum heated to 50 deg C.08 gm MgSO4 0.C's Carbol Methyl Green 1 gm Methyl green 5 gm Phenol or Carbolic 10 ml 95% ethanol 90 ml water Put phenol in mixture o 10 ml alcohol and 60 ml of water.5 gm iron alum in 100 ml of water with 14 ml glycerine.7 ml . Cool and filter. **************************************************************** Carmine. Add remaining water.17 gm CaCl2 0. BALANCED SALT SOLUTION = distilled water 100 ml NaCl 2.1M NaH2-PO4 945 ml distilled water 50 ml ballanced salt sol'n 0. Dilute filtrate eith equal partsw of 70% ethanol **************************************************************** Carnoy's Fixative 6 parts 100 % ethanol 1 part glacial acetic acid 3 parts Chloroform **************************************************************** Carnoy-LeBrun Fluid 30 ml 100% ethanol 30 ml glacial acetic acid 30 ml Chloroform aprox 10gm Mercuric Chloride Saturat mercuric chloride in the solution. **************************************************************** Carl's Solution (FAA) 170 ml 95% ethanol 60 ml Formalin 20 ml Glacial acetic acid 280 ml water Add the glacial acetic acid to the solution just before using. buffer and pour into appropriate vessels. Dissolve phenol with constant stirring. Cool and filter. Decant and filter. Put methyl green in 30 ml of water and heat.08 gm MgCl2 0.02 gm KCl 0. **************************************************************** Cerophyl Infusion for Stock Culture 4. Cool to room temp and adjust to pH 0.3 ml .1M Na2-HPO4 0. Grenacher's 100 ml distilled water 3 gm aluminum ammonium sulfate 1 gm Carmine Combine and boil for 15 min or until carmine is dissolved. Heat to 95-100 deg C for 15 min. Semicon's 1 part glacial acetic acid 1 part water Add carmine powder in excess of that which dissolves immediately. Place 1 gm Celestine Blue B in a beaker. Break up this mass coursly and pour on.46 gm .5 ml con sulfuric acid and mix with glass rod. Allow to settle for several days and do not stir when removing some stain for specimens.8 with sulfuric acid. **************************************************************** Carboxylene Melt some Carbolic acid crystals (Phenol) over a low flame (avoid breathing fumes). **************************************************************** Celestine Blue (Gray's) Dissolve 2. Pour on 0. When the effervescence has ceased the dye will be in the form of a friable mass.5 gm Cerophyl Add cerophyl to about 200 ml of boiling water and boil for 3 min. Tilt the beaker and tap to accumulate the dye in one place. Alum. Mix one part acid to three parts Xylene. Heat without boiling. with constant stirring.

Reflux all day. Add acid with swirling.5% fast green in 100 % ethanol 4.0125 gm Colchicine 25 ml water **************************************************************** . clear at room temp. or Lacmoid to make a 2% solution. tap water 3. While wet the pater will remain pink.**************************************************************** Chromate Cleaning Solution 1 120 gm Na dichromate 1000 ml tap water 1600 ml sulfuric acid Dissolve chromate in water. transfer tislsue to a vial of 5% aq solution of NaOH.rinse in water. when tissue is milky white. Place the stain with the appropriate volume of 45% acetic acid in a flask with a vertical condensor. **************************************************************** Chromate Starch 5 gm Potassium dichromate 1 gm soluable starch 100 ml water Suspend the starch and the potassium dichromate in cold water with stirring. Dilute acetic acid (4-5 %) 2. **************************************************************** Chromate Cleaning Solution 2 120 gm Potassium dichromate 600 ml water 920 ml sulfuric acid Dissolve chromate in water. 4. but when dried in the sun or near a bunsen burner. **************************************************************** Clearing Solution for leaves pg 2 of 2 4.5% crystal violet in 100 % ethanol **************************************************************** Cobalt Chloride weather paper A piece of white blotting paper is immersed in a solution containing two parts cobalt chloride to one part comon salt. **************************************************************** Cleaning solution for blood diluting pipets Pull the following solutions through the pipet in order. Allow to cool and filter. Replace with fresh NaOh when discolored 3. Acetone 5. hydrolize and remove cytoplasimic material.55 deg C. **************************************************************** Colchicine Solution 0.90 100 ml water Shake together vigorously and allow to stand. if delicate. 1% safranin in 50% ethanol 3. 0. 3. Pre treatment 1. Add acid with swirling. wash thouroughly in HOH C. Be sure to surround mixing vessel with ice bath.soak in water till material sinks. 2.20 minutes. Weigh sufficient solid Carmin or Orcein. Mount the picture on a card and hang near a window wher it will quickly respond to changes in the hygrometric state of the atmosphere. **************************************************************** Chromatographic solvent. cool. B. Be sure to surround mixing vessel with ice bath. alkaline butanol sat HOH 80 ml n. Clearing 1.extract chlorophill . otherwise heat may be used . 1% analine blue in 95% ethanol 2. glass beads or pumice will help to prevent bumping. Stainging 1. 0. Air until enclosure bounces free **************************************************************** Clearing Solution for leaves pg 1 of 2 A. A picture containing sky or water dcan be cut from a book and an inset of this prepared blotting paper made to replace the sky. 2.butanol 20 ml amonium hydroxide sp gr 0. Bring the mixture to a gentle boil while stirring.A homemade one works just as well. Dried . it turns blue. 2. This is the basis of the weather pictures sold in the shops. Fresh . **************************************************************** Chromosome stain for fruit fly salivary glands 1. Distilled water 4. Never let blood dry in the pipet 1. if not cleared in several days wash thoroughly in water and soak in 10% clorox for 10 .

Coliform test broth 10 gm Protease Peptone #3 1 gm Beef extract 5 gm Lactose 5 gm Sodium Chloride 18 mg Phenol red 1 liter water **************************************************************** Cotton blue 20 ml carbolic acid 20 ml Lactic acid 40 ml glycerine 20 ml water 0. Turk's 1 ml Glacial acetic acid 1 ml Gentian Violet 100 ml distilled water Filter before use **************************************************************** Dried Blood Spread blood on glass plate. **************************************************************** Drosophila Chromosome Stain #1 2 gm Synthetic Orcein 50 ml Glacial acetic acid 50 ml 85% Lactic acid Dissolve Orcein in hot glacial acetic acid.1 gm K2-HPO4 0.05 gm MgSO4*7HOH 10 drops 1% Ferric ammonium citrate The above should be added to 1000ml of distilled deionized water. Dry in oven or in air. Place this solution in a warming plate 32 deg C and add prewarmed water slowly with swirling until desired concentration is reached. remove from heat and add the Lactic acid. remove from heat and add the water and Lactic acid. **************************************************************** D's Dahl's Pollen Mounting Medium 16 parts 85% Lactic acid 2 parts N-Butyl alcohol 1 part 95% Ethanol 1 part Glacial Acetic Acid 1 part Glycerine **************************************************************** Desicote 5 ml Beckman Desicote 95 ml Benzene (thiophene free) **************************************************************** Dichlorophenoxyacetic acid (2-4-D) Mix dry reagent in a small amount of 95% ethanol. Drosophila Chromosome Stain #2 1 gm Synthetic Orcein 45 ml Glacial acetic acid 25 ml 85% Lactic acid 30 ml distilled water Dissolve Orcein in hot glacial acetic acid. This can be solidified by adding agar to reach a 1.5% concentration and heating to dissolve it. Filter. Filter.9 gm Crystal Violet (85% dye content) 100 mls Water **************************************************************** Cyanophycean Medium 5 gm KNO3 0.05 gm Cotton blue **************************************************************** Crystral violet 1:1000 0. **************************************************************** Dichromate Cleaning Solution 120 gm Na Dichromate 1000 ml Water 1600 ml Con Sulfuric Acid (technical grade) **************************************************************** Digestive juice (synthetic) 7-10 ml con HCl to 1 Liter Distilled water 6 gm Pepsin **************************************************************** Dilution Fluid. When dry scrape into beaker. **************************************************************** .

.A. (Carl's Solution) 170 ml 95% ethanol 60 ml Formalin 20 ml Glacial acetic acid 280 ml water Add the glacial acetic acid to the solution just before using.A. Navashin's SOLUTION A 5 gm Chromic acid 50 ml Acetic acid 320 ml distilled water SOLUTION B 200 ml formalin 175 ml distilled water Mix equal parts of each solution just before use.E's Eosin (microtechnique) 0. formalin Used to preserve color in plants **************************************************************** Formalin.02 gm NaHCO3 100 ml distilled water ** 0.5gm Eosin 100 ml 95% Ethanol **************************************************************** Erlich Solution for Indol Test 1 gm Para dimethylamino benzaldehyde 95 ml 95% ethanol 20 ml con HCl Dissolve the powder in the alcohol. Smith's 90 ml 85% isopropanol 10 ml glacial acetic acid 10 ml Formalin **************************************************************** Farmers fluid 3 parts 100% ethanol 1 part Glacial acetic acid **************************************************************** Fixative.1 gm Dextrose ** add only if requested and if requested add just before use.8 gm NaCl 0.A.A.. Neutral buffered. Transeau's 6 parts supernatant of sat CuSo4 3 parts 95% ethanol 1 part 40% aq. Botanical 115 ml Formalin 115 ml Glacial acetic acid 805 ml 95% Ethanol 805 ml Distilled water **************************************************************** F. Lavdowsky's 20 ml Formalin 100 ml Ethanol 4 ml Glacial acetic acid 80 ml distilled water **************************************************************** F..A.02 gm KCl 0.A.02 gm CaCl2 anhydrous 0.A.5 gm Na2-HPO4 **************************************************************** Frog Ringers 0.A. **************************************************************** F. Alexander's 120 ml Formalin 300 ml 95% Ethanol 20 ml Glacial acetic acid 600 ml tap water **************************************************************** F. Lillie's 100 ml formalin 900 ml distilled water 4g NaH2-PO4 6. **************************************************************** Fixative. Then add the HCl and mix well **************************************************************** F's F.A.A. .

Store in clear glass container. mold inhibitor.**************************************************************** FRUIT FLY MEDIA 37 GM BREWERS YEAST 150 GM CORN MEAL 35 GM AGAR 200 ML UNSULFURED MOLASSAS 3 GM MOLD INHIBITOR (CAROLINA BIOLOGICAL 1900 ML DISTILLED WATER Mix yeast.2 gm Sodium Sulfate 33. Turn occasionally so that all portions will be dry.45 or 70-80 % 4 ml Glacial acetic acid 20 gm Mercuric chloride 100 ml 60% ethanol 880 ml distilled water Can be heated to dissolve but let sit for a day or so to let it pcppt out.3 ml Glacial acetic acid 200 ml distilled water **************************************************************** Grenacher's Alcoholic Borax Carmine 3 gm Carmine 4 gm Borax 100 ml distilled water Boil until the carmine is dissolved (1/2 hr) or better yet . Do not use metal probes as the mercuric chloride eats metal and will cause specimen to stick to probe. Add some of the water to the corn meal to wet it and then add all the ingredients to the remaining water. C overnight with stirring and heat. **************************************************************** G's Gastric Juice or enzyme 1-2 gm pepsin 100 ml 0. WORKING DILLUTION = 1 ml of above stain to 25 ml of distilled water. While stirring constantly bring the mixture to a boil. When miced. Remove and dry on newapaper 10 .2% HCl = 6 ml con HCl in 1 liter of distilled water **************************************************************** Geimsa Stain Dissolve 0. mix and filter. Dispense in 50 ml alaquats in 1 pint milk bottles.2% HCl 0. cleaned of sand and epiphytes. making sure they are completly immersed (a weight may be added if necessary) and preserve for about 10 days. **************************************************************** Glycerine Alcohol 90 parts 70% ethanol 10 parts Glycerine **************************************************************** Glycerine Jelly 10 gm gelatin 60 ml distilled water 70 ml Glycerine 1 gm Phenol Dissolve phenol in water then stir in the glycerine. Glycerine solution may be used several times. **************************************************************** Glycerine Preservative for Kelp 3 parts Water 1 part Glycerine 10 gm/L Phenol Add kelp. To this add 33 ml of methyl alcohol. **************************************************************** Gilson's Mixture 15 ml Nitric acid sp =1.5 gm of powder in 33 ml of glycerine at 55 to 66 deg. **************************************************************** Gentian Violen Stain 1 gm Gentian Violet 20 gm Ethanol 80 ml 5% Formalin Dissolve the Gentian Violet in the alcohol then add the formalin. Transfer to a widemouth jar and allow to cool. and agar with a little 95% ethanol. add gelatin and heat until dissolved. **************************************************************** Gower's R-B-C 12.14 days.

This may take several days. Then add 100 gm of chloral Hydrate and wait until dissolved. **************************************************************** Hematoxylin.5 gm Hematoxylin 20 gm Aluminum Ammonium Sulfate 100 ml distilled water 0.0 gm Mercuric Chloride 1.5 gm (ferric ammonium sulfate) is dissolved in 100 ml of distilled water. Add the mercuric oxide with care as there is a tendency for the solution to boil up. Ferric alum (clear violet crystals) 2. **************************************************************** Holtfreter's Solution 0. They are not to be mixed. Iron 2 solutions are used. Stir and filter (takes 24 hrs). **************************************************************** H's Hagerup's Fixative 1 part propionic acid 6 parts 100% ethanol 3 parts chloroform **************************************************************** Harris' Alum Hematoxylin 0. **************************************************************** .01gm CaCl2 0.02gm NaHCO3 100 ml distilled water **************************************************************** Honey Peptone Agar 10 gm Peptone 60 gm Honey 15 gm agar 1 Liter distilled water **************************************************************** Houpt's Adhesive 1 gm of plain Knox gelatin (or reagent gelatin) is dissolved in 100 ml of water at 30 deg C.5 gm Potassium Dichromate 5.5 gm Hematoxylin 20 gm Aluminum Ammonium Sulfate 100 ml distilled water 0.0 gm Sodium sulfate 100 ml distilled water 5 ml Formalin **************************************************************** Hematoxylin. **************************************************************** Helly's Fluid 2.allow the mixture to stand at room temp until dissolved. 1.35 gm NaCl 0. After everything is dissolved add glycerine. 5 gm of Hematoxylin is dissolved in 1 liter of distilled water and 100 ml of 95% ethanol. Cool and filter. 2. Add 2 gm phenol and 15 ml of glycerine. Add the mercuric oxide with care as there is a tendency for the solution to boil up.5gm Mercuric Oxide Boil the hematoxylin and ammonium alum together in distilled water ( use a large container). **************************************************************** Hoyer's Medium 25 ml water 15 gm Gum Arabic 10 ml Glycerine 100 gm Chloral hydrate First add crystal of chloral hydrate the size of a pea to prevent fungal growth. Harris' 0.005 gm KCl 0.5gm Mercuric Oxide Boil the hematoxylin and ammonium alum together in distilled water ( use a large container). Then soak gum-water-pea of chloral hydrate for 24 hrs. Then add 70% ethanol (100 ml) and allow to stand for a day or two then filter **************************************************************** Grenacher's Alum Carmine 100 ml distilled water 3 gm aluminum ammonium sulfate 1 gm Carmine Combine and boil for 15 min or until carmine is dissolved.

Reagent to devolatalize Iodine 0. **************************************************************** . 500 ml 95% ethanol. Methyl Orange 0. Add the IAA and mix thoroughly with a glass rod and allow to cool. **************************************************************** Indol Acetic Acid in Lanolin 5 gm lanolin 2 mg IAA Warm (do not boil) the lanolin in a small screw cap vial until the lanolin just turns to liquid.5 gm Sodium thiosulfate 0. **************************************************************** Indol acetic acid solution Indol acetic acid must first be dissolved in a small amount of 95% ethanol before adding the appropriate amount of water to reach the desired concentration NOTE: IAA is unstable as a solution and bust be refrigerate Even cold it will only be good for about 36 hours.4 gm BCG.2 ml 10% CaCl2 **************************************************************** Insect Saline (Hayome) 0. **************************************************************** Iodine .2gm cresol red Thymol blue. **************************************************************** Indol Test (Kobacs' Reagent) 75 ml Amyl alcohol 25 ml con HCl 5 gm paradimethylaminobenzaldehyde Should be made up in small quantities and stored in refrigerator in clear glass stoppered bottles. 500 ml distilled water. Same procedure as above using 0. Cresol Red. pH Brom Cresol green.5 gm Methyl orange 1 liter water **************************************************************** Indicators. 1.0 gm phenolphthalein in 50 ml of 95% ethanol and 50 ml of distilled water.7% NaCl 0. **************************************************************** I's Indicator. **************************************************************** Insect Saline 0.7% NaCl in distilled water. Add distilled water in small portions to wash out the contents into a graduated cylinder.3 gm Potassoum Iodide to 1 liter distilled water **************************************************************** Iodine solution (LUGOL'S) 50 gm Iodine 100 gm KI to 1 liter distilled water Mix I and KI in a mortar.8 gm NaOH 0. 0. **************************************************************** Iodine Solution (STOCK for starch) 35 gm KI 5 gm I 500 ml distilled water for working dillution to use in starch test dilltue 1 part stock with 2 parts water. Same as above using 0. Filter.Hydrochloric acid dillutions 1 N 86 mls to volume of 1 liter 2 N 172 mls to volume of 1 liter 3 N 258 ml to volume of 1 liter **************************************************************** HYDROCHLORIC ACID DILLUTIONS 3N 258ml/L 36% **************************************************************** Hydroxide Sodium Iodide for Aquatic Analysis Add 100 grams of NaOH and 27 grams of NaI to 1 liter of water.5 gm of thymol blue Phenolphthalein.

12 gm KCl 1.6 1 gm MgSO4*7HOH 0.0 gm KNO3 1 gm Ca(NO3)2*4HOH Add each of the above to 250 ml of distilled water then combine to 1 liter adding Ca Nitrate last. **************************************************************** L's Lactophenol Mounting medium 1 part phenol crystals 1 part Lactic acid 1 part glycerine 1 part distilled water Langeron's recipie calls for 2 parts glycerine to 1 part of the other reagents. **************************************************************** Kreb's Manometer Fluid 44 gm NaBr anhydrous 0.033 at 20 deg C.25 gm KH2-PO4 0. Add 1 drop of 1% ferric chloride (freshly prepared) **************************************************************** Knops Solution (acid medium pH aprox 6. Add 1 drop of 1% ferric chloride (freshly prepared) **************************************************************** Kobacs' Reagent 75 ml Amyl alcohol 25 ml con HCl 5 gm paradimethylaminobenzaldehyde Should be made up in small quantities and stored in refrigerator in clear glass stoppered bottles.A. **************************************************************** Kymograph Shellac Solution Dillute 1 volume of shellac with 2. **************************************************************** Kovacs' Reagent . stoppered bottles.K's K.67 ml distilled water **************************************************************** Knop's solution pH 7.0 gm Ca(NO3)2*4HOH Add each of the above to 250 ml of distilled water then combine to 1 liter adding Ca Nitrate last.5ml 10% aq chromic acid 10 ml 10% aq acetic acid 0.5 volumes of 95% ethanol. **************************************************************** Lavdowsky's fixative 20 ml Formalin 100 ml Ethanol 4 ml Glacial acetic acid 80 ml distilled water **************************************************************** Litmus solution Add powdered litmus to distilled water to make a dark blue solution **************************************************************** .S.25 gm MgSO4*7HOH 0.3 gm Triton X-100 0.75 ml 95% isopropanol 105.4) 0.25 ml glacial acetic acid 263 ml sec-butyl alcohol Add sec-butyl alcohol last until solution clears **************************************************************** Karpaenchenko's fixative (Taylor's modification) 1.83 ml formalin 23.3 gm Evans blue 1 liter distilled water Density = 1.for indol test 75 ml Amyl alcohol 25 ml con HCl 5 gm Paradimethylaminobenzaldehyde Should be made up in small quantities and stored in refrig in clear glass. 158 ml crude Kerosene 473.20 gm KH2-PO4 1.A.

033.5 gm Al chloride 4. 1% Phosphotungstinic acid. 1% acid fuchsin Differentiating Moderating solution. Brodie's 23 gm NaCl 5 gm Sodium Choleate 500 ml distilled water density = 1. Pour off water add acid. Drain off water and cover with 5 volumes of 70% ethanol.2 gm NaHCO3 0. Second Staining Solution. **************************************************************** Mallory's Triple stain First Stain Solution.5 gm Analine blue 2. Add distilled water in small portions to wash out the contents into a graduated cylinder.42 gm KCl 0.5 gm KI 5 gm I 20 ml water 20 gm Chloral Hydrate **************************************************************** Methocel 2.25 gm CaCl2 2 gm dextrose 1000ml distilled water Dissolve CaCl2 in a little of the water and add it to the rest of the solution to prevent pcptation. Determine the density of the solution with a pycnometer.5-3 gm Methocel 90 ml distilled water Mic methocel with the 90 ml of Hot not boiling water.017%) **************************************************************** LUGOL'S iodine solution 50 gm Iodine 100 gm KI to 1 liter distilled water Mix I and KI in a mortar. **************************************************************** M's Maceration Fluid 1 part 10% Nitric acid 1 part 10% Chromic acid (chromic trioxide) Boil material 1 min in water. stir with glass rod and check at intervals until it begins to seperate.3 gm Triton X-100 0. Cool to 5 deg C (in refrigerator). Pour off acid. Po = 10000 Evans blue (200mg/L) or acid fuchsin are excellent dyes for the fluid.3 gm Methylene blue 30 ml 95% ethanol 70 ml Dilute KOH (0. Kreb's 44 gm NaBr anhydrous 0. other dyes may be used.0 gm CaCl2 100 ml 70% ethanol **************************************************************** Melzer's Reagent 1. It should be transparent when finished. and then at the last moment.Locke's Physiological Saline 9 gm NaCl 0.3 gm Evans blue 1 liter distilled water Density = 1. Add dextrose only if specified. **************************************************************** Mayer's Paracarmine 1 gm Carminic acid 0.033 at 20 deg C. 100 ml water 0. . but some of these tend to decompose in the manometer. put in white silk scarf or handkerchief and rinse with water for 12 hours. **************************************************************** Loeffler's methylene blue 0.0 gm Orange G 2 gm Oxalic Acid **************************************************************** Manometer Fluid. **************************************************************** Manometer Fluid.

1% N.6 gm 1-naphthylamine hydrochloride in distilled water to which 1.3 gm Methylene blue 30 ml 95% ethanol 70 ml Dilute KOH **************************************************************** Meyer's Adhesive Add to one fresh egg white. an equal quantity of glycerine and 1 gm of sodium salicylate and a crushed crystal of thymol. **************************************************************** Naphthylamine Hydrochloride Dissolve 0. **************************************************************** Nitric acid dillution 3N 195 ml/L 69% **************************************************************** . **************************************************************** Navashin's Fixative SOLUTION A 5 gm Chromic acid 50 ml Acetic acid 320 ml distilled water SOLUTION B 200 ml formalin 175 ml distilled water Mix equal parts of each solution just before use.0 ml of con HCl has been added. Lillie's 100 ml formalin 900 ml distilled water 4g NaH2-PO4 6. Loeffler's 0. B. until jelly becomes as dark as green ink.5 gm Methyl orange 1 liter water **************************************************************** Methylene blue stain 0. Thomas prefers n-butanol. then dilute with water and filter **************************************************************** Methylene Blue. Dilute to 100 ml with distilled water. 1% alpha-naphthol in 95% ethanol.5 gm Na2-HPO4 **************************************************************** Newcomers Fixative 6 parts isopropyl alcohol 3 parts propionic acid 1 part petrolium ether 1 part acetone 1 part dioxane **************************************************************** Ninhydrin Reagent 0.3 gm Methylene blue 30 ml 95% ethanol 100 ml distilled water Dissolve stain in alcohol. Store in pyrex bottle out of direct sunlight. **************************************************************** Methyl Orange Indicator 0.N-dimethyl-p-phenylenediamine HCl in water Solutions a and b are mixed in equal amounts just prior to use. **************************************************************** Nessler Reagent Dissolve 100 gm anhy7drous mercuric iodide and 70 gm anhydrous potassoum iodide in a small quantity of water and add mixture slowly to a cool solution of 160 gm of NaOH in 500 ml of distilled water.5% ninhydrin in acetone or n-bunanol. Shake wall and filter through 2-3 thicknesses of sterile cheesecloth **************************************************************** N's NADI Reagent A. Store in refrigerator. **************************************************************** Neutral Buffered Formalin.**************************************************************** Methyl Green 1 gm Methyl green 1 ml glacial acetic acid 99 ml water Dissolve satin in acid and dilute with water **************************************************************** Methyl Green Glycerine-Jelly Stain Add saturated (about 3%) solution of methyl green in 50% ethanol drop by drop to melted glycerine jelly ( a good comercial grade is satisfactory). Dilute to 1 liter with distilled water.

. Cresol Red.0 gm phenolphthalein in 50 ml of 95% ethanol and 50 ml of distilled water. Maybe kept overnight in refrigerator.2gm cresol red Thymol blue. LOCKE'S 9 gm NaCl 0. **************************************************************** P's Paradon (cellodine) 1:1 !00 % paradon and di-ethyl ether **************************************************************** Pepsin in HCl 10-2 gm Pepsin 100 ml 0.2% HCl **************************************************************** Pepsin Solution (make fresh) 1-2 gm Pepsin 100 ml water **************************************************************** Peroxides Test Add 1 ml of 10% KI to 10 ml of solution to be tested. 500 ml 95% ethanol.4 gm BCG. Add 75 ml fuming sulfuric acid (15% free SO4 Stir well and heat for 2 hours in water bath **************************************************************** Phenolphthalein 0. Incubate at body temperature Unstable.42 gm KCl 0. Dilute to a final volume of 500 ml. **************************************************************** Phenol Cotton Blue 20 ml Carbolic acid 20 ml Lactic acid 40 ml Glycerine 20 ml Water 0.O's Ortho-Toluidine Reagent (for aquatic analysis) Dissolve 4 gm ortho-toluidine in distilled water containing 15 ml of con HCl.25 gm CaCl2 2 gm dextrose 1000ml distilled water. **************************************************************** Pancreatic juice. Add dextrose only if specified.05 gm Cotton blue **************************************************************** Phenol Red Lactose Broth 10 gm Protease Peptone #3 1 gm Beef extract 5 gm Lactose 5 gm Sodium Chloride 18 mg Phenol red 1 liter water **************************************************************** Phenoldisulfonic acid Dissolve 25 gm pure white phenol crystals in 150 ml con Sulfuric acid.4 gm Baking soda 200 ml water Mix in diatomaceous earth and then filter under suction Add a few drops of chloroform. **************************************************************** pH indicators Brom Cresol green. 500 ml distilled water. artificial 40 gm Pancreatin 0.5 gm Phenolphthalein 100 ml 50% ethanol **************************************************************** Phloroglucinol. HCl Solution A. Dissolve CaCl2 in a little of the water and add it to the rest of the solution to prevent pcptation.2 gm NaHCO3 0. Same as above using 0.5 gm of thymol blue Phenolphthalein. 50 ml con HCl in 50 ml distilled water **************************************************************** Phosphoric acid dilltuion 3N 205 m/L 85% **************************************************************** Physiological saline. Filter. 0. and then at the last moment. 1. 1 gm phloroglucinol in 100 ml of 95% ethanol Solution B. If brown color results in either layer peroxides are present and the solution should be discarded with proper precautions. Same procedure as above using 0.

stir with glass rod and check at intervals until it begins to seperate.decant leaving excess CuSO4 ************************************************************** Preservative (Sutton's) 10% glycerine 3% Phenol 2% Formalin 85% Water ************************************************************** Pseudomonas Ovalic Culture Medium 1% Yeast extract 1% Beef extract 1. Filter while warm.Aquatic analysis tests for O2 20 gm K2-C2-O4*HOH dissolved in water Add 45 gm NaOH and diltue to 1 liter with water.9% NaCl 0.02% NaHCO3 0.042% 0. LOCKE'S 0. ************************************************************** Potassium Oxalate . ************************************************************** Pollen Acetylizing Mixture 9 parts Acetic Anhydride 1 part con Sulfuric Acid ************************************************************** Pollen Mounting Medium. free from contamination.24% 0.02% 0.9% 0. keep in stoppered containers.1% NaCl CaCl2 KCl NaHCO3 Dextrose (if requested) ************************************************************** Rat Ringers. ************************************************************** Prescott's Preservative for Green Plant Material 6 parts water 3 parts 95% ethanol 1 part Formalin 5 ml/100ml Glycerine Saturated with CuSO4 . put in white silk scarf or handkerchief and rinse with water for 12 hours.1% Dextrose ************************************************************** Rennin Solution 1-2 gm Rennin 100 ml water ************************************************************** . Dahl's 16 parts 85% Lactic acid 2 parts N-Butyl alcohol 1 part 95% Ethanol 1 part Glacial Acetic Acid 1 part Glycerine ************************************************************** Pollen Trapping Oil 2 parts melted white petrolatum 1 part Parafin oil 1 part butyl alcohol 1 part xylene Mix in beaker on hot blate.8% agar ************************************************************** Pyrogallic Acid 50 mg KOH 100 ml distilled water 5 gm Pyrogallol ************************************************************** R's Rat Ringers 0. Drain off water and cover with 5 volumes of 70% ethanol. Pour off acid.24% CaCl2 0.042% KCl 0.************************************************************** Plant maceration fluid 1 part 10% Nitric acid 1 part 10% Chromic acid (chromic trioxide) Boil material 1 min in water. Pour off water add acid.

After dissolved add: 95% ethanol 25 ml distilled water 25 ml Sodium acetate 1 gm Formalin 2 ml ************************************************************** Saline. Shake 2 minutes and leave in the dark for 24 hours. Cool to room temp and dilute to 100 ml by adding additional 0.1 gm Dextrose ** add only if requested and if requested add just before use.042% KCl 0. Physiological. Insect (Hayome) 0.02% CaCl2 0.24% CaCl2 0.9% NaCl 0. Insect 0. Store refrigerated. ************************************************************** SCHIFF'S REAGENT 5 gm Basic Fuchsin 1N HCl 100 ml K2-S2-O5 10 gm activated charcoal (norite) 5 gm Disolve 5 grams of basic fuchsin in 900 ml boiling distilled water.unstable 10 ml of stock brought to 500 ml with distilled water. Add dextrose only if specified.02 gm CaCl2 anhydrous 0.02% KCl 0.25 gm CaCl2 2 gm dextrose 1000ml distilled water Dissolve CaCl2 in a little of the water and add it to the rest of the solution to prevent pcptation. Cool to 50 degrees C and add 100 ml of 1N HCl cool to 25 deg and then add 10 grams of K2S2O5.2 ml 10% CaCl2 ************************************************************** Saline.02 gm NaHCO3 100 ml distilled water ** 0.7% NaCl in distilled ************************************************************** Saline. ************************************************************** Ringers for lower vertebrates 0. Working. Dilute 1:10 before using.Riboflavin Stock Dissolve 5 mg of Riboflavin in 30 ml of 0. Frog 0. Add 5 grams of fine activated charcoal. Store in a foil covered bottle in the refrigerator ************************************************************** Schoudinn's Fluid 66 ml sat aq. shake for 2 minutes and then filter through fast paper. Locke's 9 gm NaCl 0. and then at the last moment.65% NaCl 0. Filtrate should be clear. Warm in a water bath with constant rotation until dissolved.02% NaHCO3 ************************************************************** Ringers. Standard .02% NaHCO3 0.42 gm KCl 0. Mercuric chloride 33 ml 95% ethanol 5 ml glacial acetic acid Add acid just prior to use.1% Dextrose (if requested) ************************************************************** S's Safranin for plant material Dissolve 1 gm of Safranin O in 50 ml of methyl cellosolve.2 gm NaHCO3 0.02N acetic acid.8 gm NaCl 0.02 gm KCl 0. .7% NaCl 0. ************************************************************** Ringers.02N acetic acid. Rat 0.

************************************************************** Sulfuric acid dillutions 3N 168 ml/L 95% ************************************************************** Sutton's Preservative 10% Glycerine 3% Phenol 2% Formalin 85% Water ************************************************************** Sutton's preservative 10% glycerine 3% Phenol 2% Formalin 85% Water ************************************************************** T's Transeau's Fixative 6 parts supernatant of sat aq.05% Colchicine 4-6hrs sat aq Paradichlorobenzene 4 hrs sat aq alpha bromonapthalen 4 hrs ************************************************************** Starch solutions Make a thin slurry with the starch and a small amount of the water.A. Cool and filter. CuSo4 3 parts 95% ethanol 1 part 40% aq. Cool before use. ************************************************************** Smith's F. ************************************************************** Sudan IV Saturate sudan IV in absolute ethanol.5 volumes of 95% ethanol.1% final concentration works best for iodine test ************************************************************** Sudan Black B 0. Cover with a glass plate and steam for 1 hour on 2 consecutive days.************************************************************** Semicon's Carmine 1 part glacial acetic acid 1 part water Add carmine powder in excess of that which dissolves immediately. ************************************************************** Sulfanilic Acid Completely dissolve 0. Cool the solution and add 20 ml con HCl.A. Dilute to 100 ml with distilled water and mix thoroughly. 0. ************************************************************** Spindle Inhibitor in Mitosis 0.01-0. Add starch slurry while stirring bring back to a boil.5 gm calcium carbonate enough garden soil to make a layer aprox 1/2 to 3/4 inch deep and distilled water to within one inch of the top.let stand overnight and filter. Heat the rest of the water to boiling. Dilute filtrate eith equal partsw of 70% ethanol ************************************************************** Shellac solution for Kymographs Dillute 1 volume of shellac with 2. 90 ml 85% isopropanol 10 ml glacial acetic acid 10 ml Formalin ************************************************************** Soil Extract Agar 960 mls Bristols solution 40 ml soil water supnatant 15 gm agar ************************************************************** Soil water Medium In one gallon battery jar place 0.25 gm Sudan Black B 100 ml Ethylene glycol Dissolves slowly .60 gm sulfanilic acid in 70 ml of hot distilled water. Heat to 95-100 deg C for 15 min. Very viscous. formalin ************************************************************** .

************************************************************** Z's Ziehl-Nielsen's Carbolfuchsin . 100 ml water 0.5 gm Analine blue 2.1 gm Ca(NO3)2 0. Let stand all day.035 gm MgSO4 0. Mix 40 ml of this stock solution with the amount of 1N NaOH indicated below and dillute to a final volume of 100 ml using water.0 gm Orange G 2 gm Oxalic Acid ************************************************************** Tris-Maleate buffer Dissolve 29 gm of maleic acid and 30. Filter. Iron sequestrin solution (adjust to pH 6. ************************************************************** Y's Yeast in Congo Red 1 pkg dry yeast 0.7 gm Phosphotungstic acid 1 ml Glacial acetic acid 100 ml distilled water ************************************************************** Triple Stain.01N KOH = 2.61 gm Sequestrin AA 2. Mallory's First Stain Solution.00075 gm KI 0.3 gm Basic fuchsin 90% dye content 10 ml 95% ethanol 5 gm Phenol 95 ml distilled water Dissolve fuchsin in alcohol.Acid Fast 0.0 with 0. Dissolve Phenol in water and then mix solutions.0125 KH2-PO4 0. shake and let stand for 10 min. Store in refrigerator. 2% (NH4)2-HPO4 5% CaSO4.08 gm KNO3 0.3 gm Fast green FCF 0.6 gm Chromotrope 2R 0.01N HCl or 0.0025 gm Fe2-(SO4)3 0.5gm Congo red 150 ml water Add yeast to warm water and stir in congo red. ************************************************************** Turk's Diluting Fluid 1 ml Glacial acetic acid 1 ml Gentian Violet 100 ml distilled water Filter before use ************************************************************** W's Waris Solution To 1 liter of distilled water add 1 ml of the following: 10% KNO3.0016 gm H3-BO4 20 gm Sucrose Bring to 1 liter with distilled water.0015 gm ZnSO4 0.25-0. .3 gm of tris-hydroxymethyl aminomethane in 500 ml of water. 1% acid fuchsin Differentiating Moderating solution.065 gm KCl 0.0044 gm MnSO4 0. Heat to boiling and cool. Second Staining Solution.Trichrome Stain 0. 1% Phosphotungstinic acid. Add 2 g of Charcoal. 2% MgSO4*7HOH.49 gm FeSO4*7HOH 27 ml 1 N KOH 500 ml distilled water ************************************************************** White blood cell dilution fluid 1% Glacial acetic acid ************************************************************** White's Basic Medium 0.